New block-copolymer thermoassociating matrices for DNA sequencing: Effect of molecular structure on rheology and resolution

• Published in: Electrophoresis 2001-02, Vol.22 (4), p.720-728
• Main Authors: Sudor, Jan, Barbier, Valessa, Thirot, Sylvie, Godfrin, Dominique, Hourdet, Dominique, Millequant, Michelle, Blanchard, Julien, Viovy, Jean-Louis
• Format: Article
• Language: English
• Subjects: Acrylic Resins - chemical synthesis; Acrylic Resins - chemistry; Chemical Phenomena; Chemistry, Physical; DNA sequencing; Electrophoresis, Capillary - methods; Hot Temperature; Polymers - chemistry; Rheology; Sequence Analysis, DNA - methods; Solutions; Thermoassociating polymers; Viscosity; Water
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/1522-2683(200102)22:4<720::AID-ELPS720>3.0.CO;2-N

A new family of matrices for DNA sequencing by capillary electrophoresis is presented. These matrices combine easy injection with high sieving performances, due to thermal switching between a low and a high viscosity state through a modest increase in temperature (˜20°C). They are constructed from a hydrophilic polymer backbone with grafted lower critical solution temperature (LCST) side chains. The comb‐like LCST copolymers are characterized in terms of size of the polymer backbone, the size of LCST side chains and the grafting densities. The dependance of rheological behavior and electrophoretic performance of these copolymers is correlated with their microstructure. Without complete optimization, a resolution of order 0.5, corresponding to a very reasonable limit for read length with current base calling softwares, could be achieved for segments around 800 bases differing by 1 base in less than one hour in a commercial ABI 310 apparatus.